Serum estradiol was determined utilizing a radio-immunassay kit according to the method described previously (30 ). The quoted CV was less than 10% (kit information). A competitive electrochemiluminescence immunoassay (Elecsys Cortisol; Roche Diagnostics GmbH, Mannheim, Germany) was used for cortisol concentration measurement with a total CV less than 5%.
Elecsys cortisol
Manufactured by Roche
Sourced in Switzerland
The Elecsys Cortisol is a laboratory equipment product from Roche. It is used to measure the concentration of cortisol, a steroid hormone, in blood or other bodily fluids.
Automatically generated - may contain errors
6 protocols using elecsys cortisol
1
Serum Estradiol and Cortisol Assays
2
Electrochemiluminescence Cortisol Assay Protocol
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The PC levels were detected using Elecsys® Cortisol (Roche) in Cobas E411 auto analyzer. The principle of the test was electrochemiluminesence immune assay method. The sample was incubated with a cortisol specific biotinylated antibody, a ruthenylated cortisol derivative and releasing agent. The antibody binding sites were occupied either by cortisol or the ruthenylated derivative, with the proportion of each depending on the concentration of cortisol in the sample.
Streptavidin-coated micro particles were added to the reaction mixture and the immune complexes bind to the solid phase via biotin streptavidin interactions. The reaction mixture was transferred to a measuring cell and the micro particles were magnetically captured onto the surface of an electrode; unbound sample was washed away before a chemiluminescent reaction was induced by applying a voltage to the electrode. The detection limit was 0.5 nmol/L and measuring range was 0.5-1,750 nmol/L.
Streptavidin-coated micro particles were added to the reaction mixture and the immune complexes bind to the solid phase via biotin streptavidin interactions. The reaction mixture was transferred to a measuring cell and the micro particles were magnetically captured onto the surface of an electrode; unbound sample was washed away before a chemiluminescent reaction was induced by applying a voltage to the electrode. The detection limit was 0.5 nmol/L and measuring range was 0.5-1,750 nmol/L.
3
Salivary Cortisol Measurement in Runners
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Sample collections and salivary cortisol concentration measurements were performed according to a previous study [16] . The runners were not allowed to brush their teeth, chew gum, or consume any food or drink except water, 15 min before the sample collection. Saliva samples were collected using Salivette® cotton swabs (Sarstedt, Nümbrecht, Germany), centrifuged (1,500 × g) at 4°C for 10 min, then immediately stored at - 80°C until analysis. ECLIA measurements of salivary cortisol concentrations were performed using the Elecsys Cortisol on the Cobas 8000 system (Roche Diagnostics K.K, Tokyo, Japan) [16, 18] . The intra-and inter-assay coe cients of variation for salivary cortisol were 4.1% and 4.6%, respectively. The rate of change in the salivary cortisol concentration by exercise was calculated as the salivary cortisol concentration after exercise divided by the salivary cortisol concentration before exercise (%) [16] .
4
Biomarker Assessment of Inflammatory and Antioxidative Status
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We measured several blood inflammatory or antioxidative biomarkers, including cortisol, high-sensitivity C-reactive protein (Hs-CRP), total antioxidant capacity (TAC), and serum dehydroepiandrosterone sulfate (DHEA-S). Blood samples (10 mL) were collected not in a fasted state and at the same time of day at V1 and V2 to minimize the impact of circadian rhythms. The serum Hs-CRP level was determined by a turbidimetric immunoassay. 44 Plasma TAC was measured using a colorimetric assay kit (BioVision Incorporated, Milpitas, CA, USA). Serum DHEA-S levels were measured using a chemiluminescent enzyme immunometric assay. 45 Serum cortisol levels were determined using an electrochemiluminescence immunoassay kit (Elecsys Cortisol, Roche Diagnostics, Germany). All procedures were performed according to the manufacturer instructions.
5
Diurnal Salivary Cortisol Patterns
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The salivary samples were collected at 8:00 in the morning. The samples were quickly centrifuged to separate the supernatant from the cells and were then immediately frozen at −80°C until analysis. All analyses were performed in duplicate by the same researcher. Salivary cortisol was assayed using the chemiluminescence immunoassay (Elecsys Cortisol, Roche Diagnostics International AG, Rotkreuz, CH), according to manufacturer instructions. Intra- and inter- assay coefficients of variation were 4% and 6%, respectively. Salivary cortisol levels were expressed in nmol/l. Some of included (N = 75) collected salivary samples at 6 time-points: (1) on awakening; (2) at 8:00; (3) at 12:00; (4) at 15:00; (5) at 18:00 and (6) at 21:00. The participants were asked to collect the first (awakening) and the last (bed time) samples in a specific time window: Not later than 10:00 for the first and not later than midnight for the last sample.
6
Biomarker Analysis in Blood Samples
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Blood concentrations of lactate, catecholamines (including adrenaline, noradrenaline, and dopamine), cortisol, GH, IGF-1, and BDNF in the collected blood samples were analyzed. Biochemical analyses for all the indices except lactate were blindly conducted by SRL Inc. (Tokyo, Japan). Specifically, concentrations of catecholamines were measured using high-performance liquid chromatography (CA Test TOSOH; Tosoh Corporation, Tokyo, Japan), those of cortisol, GH, and IGF-1 were measured using electrochemiluminescence immunoassay (cortisol: Elecsys® Cortisol; Roche Diagnostic Systems, Basel, Switzerland, GH: Elecsys® hGH; Roche Diagnostic Systems, Basel, Switzerland, IGF-1: IGF-I IRMA DAIICHI; Fujirebio Inc., Tokyo, Japan), and the concentration of BDNF was measured using an enzyme-linked immunosorbent assay (QuantikineTM ELISA Human Free BDNF Immunoassay; R&D Systems Inc., Minneapolis, MN, USA). As mentioned above, LA was measured at each blood collection using an enzyme electrode method (Lactate Pro™2 LT-1730; ARKRAY Inc., Kyoto, Japan).
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